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Immunologically-mediated renal disease in Waldenström's macroglobulinemia.

A patient with Walderström macroglobulinemia associated with nephrotic syndrome is described. Serum cryoglobulin and rheumatoid factor were absent. Intramembranous electron-dense deposits were demonstrated in kidney biopsy material by electron microscopy. Deposits of immunoglobulin G (IgG), M (IgM) and the third component of complement (C3) were identified in kidney biopsy tissue by immunofluorescent staining methods. The serum immunoglobulins were characterized by chromatographic and immunochemical methods and showed a monocional IgM-K, IgG-K and gamma-chain piece of undefined structure. Free K- and gamma-chains were found in the urine. The IgM was not complexed to the IgG or vice versa, but the IgG was in an affregated form. Although it is not known which immunoglobulin initiated the tissue injury, IgG, IgM and complement deposits probably contributed to the renal dysfunction. The nephrotic syndrome diminished after treatemnt with chlorambucin and corticosteroids.

Animals↗

Communication between receptors for different ligands on a single cell: ligation of fibronectin receptors induces a reversible alteration in the function of complement receptors on cultured human monocytes.

Receptors for the third component of complement (C3) on cultured human monocytes (MO) bind ligand-coated particles but do not initiate phagocytosis. The function of these receptors, however, is altered dramatically after MO attach to surfaces coated with fibronectin (FN) or after MO are exposed to phorbol esters. FN and phorbol esters "activate" C3 receptors such that they promote vigorous phagocytosis. Here we show that activation of C3 receptors requires the continuous presence of FN or phorbol esters and is rapidly reversible when these stimuli are removed. Activation does not change the number or distribution of C3 receptors on the surface of MO. We conclude that the function of C3 receptors is regulated by reversible reactions that are initiated by ligation of a different class of receptors on the surface of the same cell.

Cell Communication↗

The nephropathy of experimental hepatosplenic schistosomiasis.

The glomerular lesions induced in 10 chimpanzees infected with variable numbers of Schistosoma japonicum cercariae were studied by means of light and electron microscopy and fluorescent antibody technic. Ten animals served as controls; 5 were uninfected and 5 were only lightly infected. The animals were observed for periods ranging from 3 to 17 months, and by the time of sacrifice, all had developed advanced liver fibrosis. In general, the degree of glomerular injury was related to infection intensity and degree and duration of portal liver fibrosis. Some animals had terminal BUN elevation and slight proteinuria. By light and electron microscopy, in the initial stages, only part of the glomeruli were involved and exhibited mesangial matrix expansion and mesangial cell proliferation with intracellular hyaline droplets. At later stages, a larger number of glomeruli were affected and exhibited diffuse hypercellularity, glomerular basement thickening, mesangial sclerosis and less often, focal necrosis, crescent formation, synechiae and global hyalinization. In addition, there were discrete electron-dense deposits localized in the mesangial area in some glomeruli. Immunofluorescent studies utilizing antisera to chimpanzee gamma-globulin and complement (C3) and to human properdin disclosed only faint deposits of C3, apparently in mesangial areas. The association of hepatosplenic schistosomiasis and nephropathy, the possible role of schistosomal antigen and the mechanism(s) of such glomerular injuries are reviewed and compared with the disease in humans and other host species infected with Schistosoma.

Animals↗

AT1-receptor antagonists abolish glomerular MCP-1 expression in a model of mesangial proliferative glomerulonephritis.

BACKGROUND: Glomerular accumulation of macrophages/monocytes (M/M) is a typical early feature in the course of anti-thymocyte serum (ATS)-induced nephritis. We have previously shown that glomerular synthesis and expression of monocyte-chemoattractant protein-1 (MCP-1) occurs before influx of M/M and a neutralizing anti-MCP-1 antibody reduced this cell infiltrate by one third. The present study was undertaken to test the effect of two angiotensin II type 1 (AT1) receptor antagonists, losartan and irbesartan, on ATS-stimulated MCP-1 expression as well as glomerular influx of M/M. METHODS: Treatment of rats with either losartan or irbesartan was started 24 h before administration of ATS. After 24 h, MCP-1 mRNA expression was evaluated by RT-PCR and Northern blots. MCP-1 protein was determined by Western blots and chemotactic factors released from isolated glomeruli were measured by chemotactic assay. Kidney sections were stained for rabbit IgG, complement C3, and M/M (ED1 antigen). RESULTS: Both AT1-receptor antagonists caused a significant, but not total reduction in MCP-1 mRNA and protein expression 24 h after injection of ATS. Treatment with losartan or irbesartan also reduced the chemotactic activity of isolated glomeruli from nephritic animals. Quantification of ED1-positive cells revealed that losartan as well as irbesartan reduced glomerular M/M invagination in nephritic rats by approximately 30-50%. However, treatment with AT1-receptor antagonists did not influence binding of ATS to mesangial cells and subsequent complement activation indicating that the attenuated MCP-1 expression is not due to differences in delivery and binding of ATS to mesangial cells. CONCLUSION: Our data indicate that short-term antagonism of AT1 receptors abolished the early glomerular MCP-1 expression and M/M influx. These results indicate that angiotensin II may exert immunomodulatory effects in vivo and adds a new mechanism showing how this vasopeptide may be involved in the pathogenesis of renal diseases.

Angiotensin Receptor Antagonists↗

[Blood levels of immunoglobulins G,A,M, circulating immune complexes, complement and proteins in patients on intermittent peritoneal dialysis].

The study involved 17 patients on IPD. Blood serum levels of IgG, IgA, IgM, circulating immune complexes, complement and proteins were determinated at the beginning of therapy, after 3, 6, 12 months on IPD and after 1 year on hemodialysis. The frequency of peritonitis was noted during this time. Peritonitis was the most frequent during first 3 months on IPD. No differences in blood serum levels of IgG, IgA, IgM, in the specific periods of IPD were noted. A significant increase in blood serum circulating immune complexes in patients on IPD and hemodialysis compared to the control group was found. A significant decrease in blood serum of C3 complement in patients on IPD and hemodialysis in comparison with the controls were found. A significant decrease in blood serum proteins at the beginning of IPD and after 3 months IPD in comparison with proteins concentration in patients on hemodialysis was observed.

Adolescent↗

Immunologic subsets in B cell lymphomas defined by surface immunoglobulin isotype and complement receptor--their relationship to survival.

Cell surface marker profiles were studied on cell suspensions from monoclonal B cell lymphomas. Surface immunoglobulin (sIg) was examined in 178 cases, whereas combined data with sIg and receptors for complement (C3) were available in 99/178 cases. The results showed that B cell lymphomas can be divided into distinct immunological subsets according to surface marker expression. Whereas some histologic subgroups (diffuse centroblastic/centrocytic, centrocytic, immunocytoma) (Kiel classification) consisted of few immunological subtypes, others were more heterogeneous (follicular centroblastic/centrocytic, centroblastic, lymphocytic). By combining immunological and histological subgroups, more than 40 phenotypes could be identified; this diversity most likely reflects, the heterogeneity of the B cell compartment. As part of the same study the prognostic significance of cell marker phenotypes was examined. Survival analysis undertaken on 149/178 patients did not uncover any significant relationship between type of heavy or light chain expression, C3 receptor expression and clinical outcome. Our data do not confirm recent findings that the type of immunological phenotype may be of prognostic significance.

Aged↗

[Sweet syndrome with immune complex vasculitis in a child].

We report on a rare case of Sweet's syndrome (SS) in a 7-year-old boy showing typical symptoms such as high fever, neutrophil leucocytosis in peripheral blood, elevated blood sedimentation rate, and arthralgia. The cutaneous lesions were painful erythematous, raised plaques on both limbs and the face. Histology confirmed the clinical diagnosis, including remarkable signs of cutaneous small vessel vasculitis, which was verified by direct immunofluorescence for IgM and complement (C3) in vessel walls as well as by the presence of circulating immune complexes in the blood. By means of monoclonal antibodies, T-lymphocytes and macrophages could be identified in the perivascular infiltrates in the delayed phase of SS. According to these immunological findings, we discuss the pathomechanism and the therapeutical benefit of inhibitors of cyclooxygenase.

Capillaries↗

Impaired expression of erythrocyte glycosyl-phosphatidylinositol-anchored membrane CD59 in patients with psoriatic arthritis. Relation to terminal complement pathway activation.

OBJECTIVE: Complement-mediated injury is regulated by many factors; among these CD59 has been identified as a widely distributed glycoprotein that inhibits membrane C5b-9 (terminal complement component) formation. The aim of the study was to assess erythrocyte CD59 expression in patients with psoriatic arthritis in order to understand the role of CD59 in the pathogenesis. METHODS: Washed erythrocytes from 50 patients with psoriatic arthritis, 8 with cutaneous psoriasis and 24 healthy subjects were incubated with monoclonal anti-CD59 antibody followed by a second FITC conjugated antibody and fluorescence intensity analysed by FAC-Scan flow cytometer to assess their CD59 membrane expression. SC5b-9 levels were measured in the plasma by ELISA and results compared with CD59 values. Immune complexes, complement C3 and C4 and rheumatoid factor were also determined. RESULTS: Impaired expression of erythrocyte membrane-anchored CD59 was found in patients with psoriatic arthritis; the lowest levels were seen in active patients (p < 0.01). Increased SC5b-9 was seen in the plasma of patients with active disease. An inverse correlation was also found between plasma C5b-9 and the CD59 expression levels (r = -0.81, p < 0.001). CONCLUSION: The low CD59 expression on erythrocytes from patients with psoriatic arthritis may be an index of a low tissue CD59 expression. This impairment could facilitate the activation of complement pathway and increase the risk for arthritis. Membrane attack complex formation in deficient membrane bound CD59 may also exacerbate synovial cell injury and inflammation.

Adult↗

Immunopathogenesis of canine aural haematoma.

Data are presented from 15 dogs with aural haematoma. The series included six Labrador retrievers and four golden retrievers and the mean age was 8.0 +/- 3.02 years. Five dogs had evidence of pruritic skin disease and five further cases had other concurrent disease. Haematology and serum biochemistry were normal in 12 and 13 of the 15 dogs, respectively. All dogs were Coombs' negative and serum antinuclear antibody had negative or low titres in all the 11 cases tested. Histopathological examination of biopsies from the affected ears revealed variable degrees of erosion of auricular cartilage with fibrovascular granulation tissue filling the cartilage defects. There was minimal perichondral inflammation. The biopsies were studied by immunohistochemistry for deposition of immunoglobulin G (IgG), immunoglobulin M (IgM) and complement C3. In one dog there was basement membrane zone deposition of IgG and in another there was focal interepithelial deposition of both IgG and IgM. The findings of this study do not support an autoimmune pathogenesis for canine aural haematoma, but suggest that an early immunological event may underlie the observed cartilage erosion.

Animals↗

Role of cell surface receptors in the regulation of intracellular killing of bacteria by murine peritoneal exudate neutrophils.

The role of the Fc and third component of complement (C3) receptors on mouse neutrophils in the control of killing of Proteus mirabilis, opsonized in normal mouse serum (NMS) or heated immune mouse serum (HIMS), was studied. The events following incubation of neutrophils with P. mirabilis and the events associated with bacterial killing were assayed. The respiratory burst was quantified by chemiluminescence (CL). Levels of leukocyte-associated bacteria were determined after a 20-min ingestion period as a measure of phagocytosis. Bacterial killing was measured while ingestion was allowed to continue or as a discrete process when extracellular, noningested bacteria had been removed and neutrophils with intracellular bacteria were incubated in the presence of serum. Modification of these responses in the presence of three monoclonal antibodies (MAb), NIMP-R10 and M1/70, which bind to different epitopes of the mouse C3 receptor, and 2.4G2, which binds to the mouse Fc receptor, was investigated. MAb to the C3, but not to the Fc, receptors reduced CL, ingestion, and intracellular killing of NMS-opsonized P. mirabilis. MAb to the Fc receptor diminished CL to and reduced the rate of ingestion of HIMS-opsonized bacteria. The two MAb to the C3 receptor each produced a similar inhibition of ingestion and intracellular killing of HIMS-opsonized bacteria, but they only partially blocked CL. A range of MAb preparations reactive with other murine antigens did not inhibit these events, either with NMS- or HIMS-opsonized P. mirabilis. The results suggest that C3 receptors on mouse neutrophils played a predominant role in regulation of the killing of P. mirabilis. Similar results were found for Staphylococcus aureus. C3 receptors were necessary for maximal expression of all functions culminating in bacterial kill. That MAb to the C3 receptor inhibited phagocytosis of HIMS-opsonized bacteria in similar fashion to the effect of MAb to the Fc receptor and in contrast to the lack of effect of control MAb may reflect steric hindrance of the Fc receptor by MAb binding to the C3 receptor, or it may reflect that the receptors are linked in murine neutrophils as they are in human neutrophils.

Animals↗

The immunocompetence of children with congenital heart disease.

We studied the immunocompetence of 18 children with conotruncal malformations (13 with tetralogy of Fallot, 5 with truncus arteriosus) and 22 children with cardiac shunt lesions. There were reduced total T cell percentages and T helper cells in the conotruncal group but no T cell abnormality in the shunt group. Also, 7 of the 18 cases in the conotruncal group had facial dysmorphism reminiscent of the Di George syndrome. These results suggest that patients with conotruncal malformations fall into the wide spectrum of the Di George syndrome. There was some humoral deficiency in both groups with reduced levels of immunoglobulins IgG and IgA and low levels of complement C3 and C4. The clinical records showed a high frequency of infections. Hospital admissions for these episodes had occurred in 61% of the conotruncal group and 32% of the shunt group. Thus, there is an increased susceptibility to infection in children with congenital heart disease, and the predilection to infection has an immunological basis.

B-Lymphocytes↗

An immunoradiometric assay for erythrocyte complement (C3b) receptor activity applied to a pediatric population with connective tissue disease.

The number of receptors for complement component C3b per erythrocyte reportedly is decreased in over half of adults with systemic lupus erythematosus. We have devised an immunoradiometric assay for C3b receptor (CR1) on erythrocytes, with which one can assess CR1 saturation due to in vivo binding of immune complexes or activated complement fragments (C3b). Using this assay, we examined binding by CR1 in normal adults and newborns, in lupus and juvenile rheumatoid arthritis patients, and in a population of patients with various general medical problems, including other connective tissue diseases. Binding by CR1 was decreased in eight of 15 SLE patients, four of 25 juvenile rheumatoid arthritis patients, and one of 14 patients with other diseases. We found no significant correlation between CR1 binding and either C1q binding, antinuclear antibody titer, results for complement C3 and C4, or the presence of renal disease. Using this assay, we were also able to show that the observed reduction in CR1 binding was not ascribable to prior saturation of CR1 or to blocking antibody against CR1. The assay is precise and easy enough for routine application.

Adult↗

Diagnostic relevance of fibronectin in cryoprecipitates.

Fibronectin is a known component of plasma cryoprecipitates. It is seen in cryoglobulins from patients with monoclonal gammopathies and also from rheumatoid arthritis, and patients with systemic lupus erythematosus and other connective tissue diseases. We evaluated the clinical relevance of measures of cryoprecipitable fibronectin from the sera of 88 patients with rheumatic diseases and 27 healthy controls. There were 28 patients with rheumatoid arthritis, 19 with systemic vasculitis, 5 with cutaneous vasculitis, and 36 with a systemic connective tissue disorder. We measured total and cryoprecipitable fibronectin and for comparison immunoglobulins G, A, and M and complement C3 and C4. Cryoprecipitable fibronectin was detected in 33% control sera and 42% patient sera. The mean levels were higher in the sera of patients in all diagnostic groups. The highest levels were seen in rheumatoid patients with systemic disease, systemic vasculitis, and connective tissue diseases. The presence of cryoprecipitable fibronectin was related to the clinical activity of systemic vasculitis; none of the 6 patients with clinically inactive vasculitis had detectable fibronectin in their cryoprecipitates; but it was seen in 7 to 13 cases with active vasculitis. There were only weak relationships between cryoprecipitable fibronectin and immunoglobulin and complement levels in cryoprecipitates. We conclude that routinely measuring fibronectin levels in cryoprecipitates is generally of doubtful diagnostic value. However, it appears to be a useful marker of the clinical activity of systemic vasculitis and we recommend its use in the laboratory assessment of vasculitis.

Adult↗

C1q governs deposition of circulating immune complexes and leukocyte Fcgamma receptors mediate subsequent neutrophil recruitment.

Inflammation induced by circulating immunoglobulin G-immune complexes (ICs) characterizes many immune-mediated diseases. In this work, the molecular requirements for the deposition of circulating ICs and subsequent acute leukocyte recruitment in mice were elucidated. We show that after intravenous injection, preformed soluble ICs are rapidly deposited in the postcapillary venules of the cremaster microcirculation, secondary to increased vascular permeability. This deposition is dependent on complement C1q. IC deposition is associated with leukocyte recruitment. Leukocyte rolling, which is mediated by P-selectin in the exteriorized cremaster muscle, is not further increased in response to ICs. In contrast, leukocyte rolling velocity is significantly decreased and leukocyte adhesion is significantly increased in the presence of ICs. The IC-mediated slow leukocyte rolling velocity and subsequent adhesion and emigration are dependent on Fcgamma receptors (FcgammaRs), particularly FcgammaRIII, with complement C3 and C5 having no detectable role. These studies suggest a regulatory mechanism of IC deposition and leukocyte trafficking in IC-mediated inflammation requiring C1q and FcgammaRs in sequential, noninteracting roles.

Animals↗

The adipsin-acylation stimulating protein system and regulation of intracellular triglyceride synthesis.

We have previously characterized an activity from human plasma that markedly stimulates triglyceride synthesis in cultured human skin fibroblasts and human adipocytes. Based on its in vitro activity we named the active component acylation stimulating protein (ASP). The molecular identity of the active serum component has now been determined. NH2-terminal sequence analysis, ion spray ionization mass spectroscopy, and amino acid composition analysis all indicate that the active purified protein is a fragment of the third component of plasma complement, C3a-desArg. As well, reconstitution experiments with complement factors B, D, and complement C3, the components necessary to generate C3a, have confirmed the identity of ASP as C3a. ASP appears to be the final effector molecule generated by a novel regulatory system that modulates the rate of triglyceride synthesis in adipocytes.

Amino Acid Sequence↗

[Preservation of fresh homograft using TC199 solution in rat].

The surgical replacement of human vessels using fresh allografts (homograft) has been reported as useful means. Nevertheless, little is known about preservation effect and immunological consequences using TC199 solution. In this study, defined inbred strains of rats were used. The aorta was extracted from Buffalo rats and preserved for 1, 2, and 3 weeks in TC199 solution at 4 degrees C. The preserved graft was then implanted into the Buffalo rats heterotopically, in the abdominal aorta, in the autotransplant group (AUTO) and into the Lewis rat in the allotransplant group (ALLO). The graft patency and histology including immunohistochemical stain (IgG, IgM, and C3 complement) were assessed at 2 weeks after transplantation. In AUTO, the patency was 100% with grafts 1 and 2 weeks' preservation, and 80% with 3 weeks' preservation. In the ALLO, patency was 100% with grafts with one week's preservation, 80% in those with 2 weeks' preservation and 0% those with in 3 weeks' preservation. Histology revealed that fibroblasts and endothelial cells were significantly deteriorated in 3 weeks' preservation in auto transplantation, however, they were well-maintained in 2 weeks' preservation. In the allo-transplantation group, histological deterioration began after 2 weeks preservation. Immunohistochemical staining showed significant deposits of IgG, IgM and C3 in ALLO. Significant infiltration of antibodies was seen in deteriorated tissue due to prolongation of the preservation periods. These results suggested that freshly preserved allograft was antigenic and 2 weeks preservation using TC199 solution was possible in the rat model.

Animals↗

Localization of immunoglobulins and complement by the peroxidase antiperoxidase method in autoimmune and non-autoimmune canine dermatopathies.

Formalin-fixed, paraffin embedded skin biopsy specimens from 44 dogs with various dermatopathies were examined for the presence of immunoglobulins (IgG, IgM, IgA) and complement (C3) by the peroxidase antiperoxidase method (PAP). Final diagnoses of the skin diseases were determined by clinical findings, fungal and bacterial cultures, skin scrapings, serum endocrinologic tests, and histologic features of cutaneous biopsies. The dermatopathies included examples of pyoderma (folliculitis/furunculosis), demodecosis, sarcoptic mange, dermatophytosis, endocrine dermatopathy, and autoimmune skin disease (AISD). In the latter category, 7 cases of pemphigus foliaceus, 1 pemphigus vulgaris, 4 discoid lupus erythematosus (DLE) and 4 examples of indeterminate AISD were examined. Immunoglobulins, C3, or both, were localized in tissue sections from AISD (15/16), pyoderma (7/11), demodecosis (4/8) sarcoptic mange (3/3), and dermatomycosis (2/3). Endocrine dermatopathy biopsies were consistently negative for Ig and C3 (0/3). The pattern of localization was most often intercellular (31/44 positive biopsies) with basement membrane immunoreactivity in 4 cases of DLE, and 1 case of indeterminate AISD. There was no consistent correlation between histologic features of hematoxylin and eosin-stained biopsies and the presence of Ig and C3. Clinical outcome was similar in both Ig and C3 positive and negative cases of non-AISD dermatitis. The high percentage (95%) of positive results in AISD biopsies indicates the usefulness and sensitivity of the PAP method for the localization of Ig and C3. Because of the high percentage of Ig localization in pyoderma (73%), biopsy specimens with extensive inflammatory skin disease are not suitable for diagnosis of AISD. The PAP method is useful in the diagnosis of AISD in biopsy specimens with minimal inflammation. Caution is warranted in the interpretation of immunoreactivity independent of clinical and histologic information.

Animals↗

Follicular mucinosis: a detailed morphologic and immunopathologic study.

Two patients with the benign type of follicular mucinosis (FM) are presented. Their clinical features and course were characteristic for this subgroup of FM. Light and electron microscopy, direct immunofluorescence, and immunoperoxidase cell marker studies were undertaken to characterize the nature of the disease process. Light microscopy confirmed the follicular outer root sheath and sebaceous gland epithelial degenerative changes. The infiltrating inflammatory cells were morphologically benign. Electron microscopy detailed the cellular associations in the areas of degenerative change. Disattached keratinocytes were closely apposed to significant numbers of macrophages and Langerhans cells. Direct immunofluorescence studies demonstrated primarily complement (C3) and fibrinogen/fibrin in areas of reticular degeneration. Immunoperoxidase studies revealed large numbers of T cells and macrophages and a striking increase in the number of Langerhans cells in the affected follicular epithelium. The findings suggest that cell-mediated immune mechanisms may play a role in the pathogenesis of this disorder.

Adult↗

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